Gene Conversion Diversification of Autonomous Heavy Chain Antibodies
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Solution Overview
Problem
Existing technologies face challenges in efficiently diversifying heavy chain-only antibodies using the immune system of a host animal.
Innovation Solution
A transgenic animal is developed with a functional immunoglobulin heavy chain gene and operably linked pseudogenes that facilitate gene conversion, allowing for the diversification of autonomous heavy chain-only antibodies by mutating the nucleic acid encoding the variable domain.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If heavy chain-only antibodies are produced without light chains, then autonomous heavy chain variable domains can fold and bind to antigens independently, but the diversity of antibodies cannot be efficiently diversified using the immune system of the host animal
Solution Approach 1:
The patent introduces pseudogenes upstream or downstream of the functional heavy chain gene that are pre-configured to donate diverse nucleotide sequences through gene conversion. This preliminary arrangement of genetic elements enables the immune system to automatically diversify antibodies through natural gene conversion mechanisms during B cell development, resolving the contradiction between autonomous functionality and diversification efficiency
Solution Approach 2:
The patent uses pseudogenes as intermediary elements that mediate sequence transfer to the functional heavy chain gene via gene conversion. These pseudogenes serve as a reservoir of diverse sequences that can be converted into the functional gene, enabling efficient antibody diversification without requiring light chain pairing, thus maintaining autonomous heavy chain functionality while achieving diverse antibody repertoires
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The transgenic animal produces diversified antibodies with optimized autonomous heavy chain variable domains, maintaining functional properties through gene conversion and somatic hypermutation repair.
Implementation Method 1
a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the AHC variable domain of (a)
Implementation Method 2
the autonomous nature of the heavy chain encoded by the functional gene may be preserved by gene conversion events and any de-camelizing amino acid substitutions that occur as a result of somatic hypermutation should be repaired by gene conversion
Data Source
AI summary
This disclosure provides, among other things, a transgenic animal that uses gene conversion for antibody diversification, comprising B cells in which the endogenous immunoglobulin heavy chain locus comprises: (a) a functional immunoglobulin heavy chain gene comprising a nucleic acid encoding an autonomous heavy chain (AHC) variable domain; and (b) a plurality of pseudogenes that are operably linked to said functional immunoglobulin heavy chain gene and that donate, by gene conversion, nucleotide sequence to the nucleic acid encoding the AHC variable domain of (a), wherein the pseudogenes are upstream or downstream of the functional immunoglobulin heavy chain gene.